Functional impact of IgA nephropathy-associated selectin gene haplotype on leukocyte–endothelial interaction
Functional impact of IgA nephropathy-associated selectin gene haplotype on leukocyte–endothelial interaction
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DOI：
10.1007/s00251-006-0120-7
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发表时间：
2006-04
期刊：
影响因子：
3.2
通讯作者：
T. Takei;M. Hiraoka;K. Nitta;K. Uchida;Michiyo Deushi;Tao Yu;N. Nitta;K. Tsuchiya;W. Yumura;H. Nihei;Yusuke Nakamura;M. Yoshida
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文献类型：
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作者：
T. Takei;M. Hiraoka;K. Nitta;K. Uchida;Michiyo Deushi;Tao Yu;N. Nitta;K. Tsuchiya;W. Yumura;H. Nihei;Yusuke Nakamura;M. Yoshida
此前，我们在选择素基因中发现了与免疫球蛋白A （IgA）肾病相关的单核苷酸多态性（snp），其中L选择素的启动子区为712C>T(P238S)， L选择素的启动子区为-642A >G， E选择素的1402C>T（H468Y）。有趣的是，这些snp几乎处于完全的连锁不平衡状态，因此构建了两个单倍型，即疾病相关的TGT和野生型（Wt） CAC。为了研究TGT单倍型的功能意义，我们建立了一个表达p238s - l -选择素变体（CHO- varl）的稳定CHO转染物和一个含有h468y - e -选择素变体（Ad-varE）的重组腺病毒载体，并与它们的Wt对应物进行了比较。在流动条件下，CHO-varL对il -1β活化的HUVEC单层的粘附能力明显低于CHO-wtL。此外，含有l -选择素变体（luc-varL）启动子区域的荧光素酶报告结构，与Wt （luc-wtL）相比，表现出明显较低的转录活性。这些结果表明，L选择素在疾病相关单倍型中的粘附相互作用和表达水平显著降低，表明这些snp在炎症性疾病（包括IgA肾病）的发病机制中具有潜在作用。
Previously, we discovered single-nucleotide polymorphisms (SNPs) associated with Immunoglobulin A (IgA) nephropathy in selectin genes, which were 712C>T(P238S) in L selectin, –642A>G in the promoter region of L selectin, and 1402C>T(H468Y) in E selectin. Interestingly, these SNPs were in nearly complete linkage disequilibrium, thus two haplotypes, disease-associated TGT and wild-type (Wt) CAC, were constructed. To investigate the functional significance of TGT haplotype, a stable CHO transfectant expressing a P238S-L-selectin variant (CHO-varL) and a recombinant adenovirus vector containing an H468Y-E-selectin variant (Ad-varE) were established and compared to their Wt counterparts. Under flow, CHO-varL exhibited significantly less adhesion over IL-1β-activated HUVEC monolayers compared to CHO-wtL. Furthermore, a luciferase reporter construct, containing a promoter region of the L-selectin variant (luc-varL), exhibited significantly less transcription activity compared to Wt (luc-wtL). These results suggest that the adhesive interactions and expression level of L selectin in disease-associated haplotypes are significantly compromised, indicating a potential role of these SNPs in the pathogenesis of inflammatory diseases, including IgA nephropathy.